Pierson, Barbara J. (ORCID: 0000-0001-8233-874X) Gravley, Megan C. (ORCID: 0000-0002-4947-0236) Sage, G. Kevin (ORCID: 0000-0003-1431-2286) Talbot, Sandra L. (ORCID: 0000-0002-3312-7214) 20230816 tabular digital data Anchorage, Alaska U.S. Geological Survey, Alaska Science Center Suggested Citation: Pierson, B.J., Gravley, M.C., Sage, G.K., Talbot, S.L., 2023, DNA microsatellite and sex identification marker screening for Mexican Spotted Owl (Strix occidentalis lucida): U.S. Geological Survey data release, https://doi.org/10.5066/P9KEWTRX https://doi.org/10.5066/P9KEWTRX Two tables of data are included in this data release. The first table provides screening results for 32 previously described and/or published microsatellites and 1 sexing marker, primer redesigns for use in Mexican Spotted Owls in this data release, and size of markers in Mexican Spotted Owls. The second table provides microsatellite and sexing marker results for 10 tissue and 98 feather samples for 11 microsatellite markers identified as variable in initial marker screenings. The Mexican Spotted Owl (Strix occidentalis lucida) was listed as threatened by the U.S. Fish and Wildlife Service in 1993. The National Park Service requested assistance with developing genetic markers to identify individual owls from feathers and other materials. Genetic IDs or fingerprints were to then be used in subsequent analyses of habitat occupancy, dispersal rates, and gene flow among NPS units. 2017 2021 observed Complete None planned From Grand Canyon National Park in the west, Kaibab National Forest in the north (including most in Walnut Canyon National Monument) to the Tonto National Forest in the south to Los Alamos, New Mexico in the east. -113.999 -106.245 35.593 33.166 USGS Metadata Identifier USGS:ASC551 ISO 19115 Topic Category Biota Environment NASA GCMD Earth Science Keyword Thesaurus Animals/Vertebrates Birds USGS CSA Biocomplexity Thesaurus Birds of prey Threatened species Breeding sites Capture-recapture studies USGS Thesaurus Wildlife Terrestrial ecosystems Sexing (plants & animals) Genetics Genetic diversity Polymerase chain reaction NIH MeSH Thesaurus DNA DNA Primers Alleles Genetic markers Microsatellite Repeats Genotype None Raptors Mexican Spotted Owl Strix occidentalis lucida USGS Geographic Names Information System (GNIS) Arizona Walnut Canyon National Monument Apache-Sitgreaves National Forest Grand Canyon National Park Coconino National Forest Tonto National Forest Kaibab National Forest New Mexico Los Alamos None Mexican Spotted Owl Strix occidentalis lucida ITIS Integrated Taxonomic Information System Unknown online database online ITIS-North America Taxonomic details retrieved July 26, 2023, from the Integrated Taxonomic Information System online database https://www.itis.gov https://doi.org/10.5066/F7KH0KBK Species were identified by skilled observers in the field based on general appearance, plumage, behavior, and vocalization. Kingdom Animalia Subkingdom Bilateria Infrakingdom Deuterostomia Phylum Chordata Subphylum Vertebrata Infraphylum Gnathostomata Superclass Tetrapoda Class Aves Order Strigiformes Family Strigidae Subfamily Striginae Genus Strix Species Strix occidentalis Spotted Owl TSN: 177925 Subspecies Strix occidentalis lucida Mexican Spotted Owl TSN: 177928 No access constraints. No use constraints. We request that the suggested citation of this USGS data release be included in any publications that reference or utilize these data. U.S. Geological Survey, Alaska Science Center Mailing and Physical

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Anchorage Alaska 99508 USA 907-786-7000 gs-ak_asc_datamanagers@usgs.gov National Park Service, Walnut Canyon National Monument, 6082 Sunset Crater Road, Flagstaff, AZ 86004 Individual sample ID's, collection dates, and sampling locations of individual samples provided by collectors were cross-checked and verified to match the accompanying worksheet. For quality control purposes, DNA from 9 feathers was extracted, amplified, and run in duplicate. We observed sterile techniques in the handling of all DNA, and all procedures were performed with positive and negative controls to provide evidence of replication without contamination. Poor DNA quantity and quality were anticipated with these opportunistically-gathered feather samples and some results were observed that are consistent with this expectation and noted in individual attributes as WK for results that were observed but too weak to score with confidence, NP for no scorable product observed, or AD for samples that were homozygous, but consistent with possible allelic dropout due to low template DNA quality/quantity. Additionally, the primers used to determine sex were redesigned from Henke (2005) after sequencing a portion of the CHD gene for Mexican Spotted Owl. The new primers should amplify a product approximately 210 bp in size, but were not systematically re-sized using an M13 ladder. Reported sizes in the table "mexicanSpottedOwl_microsat_pierson.csv" are based on sizes determined with the larger Henke (2005) fragment (387/393 bp). For microsatellite loci, reported allele sizes were reviewed to confirm that they conform to expected distributions (e.g. dinucleotide repeats with 1 base pair differences were reviewed on gel images by laboratory personnel, gaps in allele sizes were reviewed, and single occurrences of an allele size were scrutinized). Nine of 99 samples were successfully re-extracted and run in duplicate with concordant results. Many samples were run multiple times for multiple loci and notations for suspected allelic drop out, false alleles, consistently weak or unscorable products are noted in the accompanying table. Data omissions for particular entities are explained in the Entity and Attribute definitions. Briefly: Collection dates are provided when available, but were not available for all samples provided to the laboratory. Blank data indicates no data were collected or not provided with the sample for a particular attribute. Henke, A.L. 2005 publication online San Jose State University Henke, AL., 2005. Spotted owl (Strix occidentalis) microsatellite variation in California. Master's Thesis, San Jose State University, https://doi.org/10.31979/etd.cjpz-p25j https://doi.org/10.31979/etd.cjpz-p25j publication 2005 publication date Henke 2005 Spotted Owl-specific CHD primers that amplify a 387-393 bp size fragment used to redesign primers to amplify a shorter fragment in this study. Redesigned primers are provided in the screen table accompanying this data release. (Henke 2005 indicates his primers were based on Boreal Owl (Aegolius funereus) CHD sequences in Fridolfsson and Ellegren 2000 (GenBank AF181824, AF181825). Hsu, Y-C Severinghaus, L.L. Lin, Y-S Li, S-H 2003 journal article Molecular Ecology Notes 3(4):595-597 online Wiley Hsu, Y-C, Severinghaus, L.L., Lin, Y-S, Li, S-H, 2003. Isolation and characterization of microsatellite DNA markers from the Lanyu scops owl (Otus elegans botelensis). Molecular Ecology Notes 3(4):595-597, https://doi.org/10.1046/j.1471-8286.2003.00523.x https://doi.org/10.1046/j.1471-8286.2003.00523.x publication 2003 publication date Hsu et al. 2003 Primers for Oe3-7-1, Oe45, Oe54, Oe128-1, and Oe2-57-1 were screened for variability in Mexican Spotted Owls. Oe3-7, Oe128, and Oe2-57 were redesigned for optimization attempts. Oe128-1 is retained as an informative marker in this data release (with the "-1" indicating primers were redesigned and provided in the screen table accompanying this data release). Hull, J.M. Keane, J.J. Tell, L.A. Ernest, H.B. 2008 journal article Conservation Genetics 9(5):1357-1361 online Springer Hull, J.M., Keane, J.J., Tell, L.A., Ernest, H.B., 2008. Development of 37 microsatellite loci for the great gray owl (Strix nebulosa) and other Strix spp.owls. Conservation Genetics 9(5):1357-1361, https://doi.org/10.1007/s10592-007-9489-6 https://doi.org/10.1007/s10592-007-9489-6 publication 2008 publication date Hull et al. 2008 Primers for SneA012, SneA001w, SneA004, SneD105, SneD116, SneD123, SneD202w, SneD224, SneA127w, SneA318, and SneA338 were screened for variability in Mexican Spotted Owls. SneA001w-1, SneD105-1, SneD116-1, SneD202w-1, and SneA338-1 were redesigned for optimization attempts. Ultimately, SneD105-1, SneD202w-1, and SneA127w were retained as informative markers in this data release (with the "-1" indicating primers were redesigned and provided in the screen table accompanying this data release). Isaksson, M. Tegelstrom, H. 2002 journal article Molecular Ecology Notes 2(2):91-93 online Wiley Isaksson, M., Tegelstrom, H., 2002. Characterization of polymorphic microsatellite markers in a captive population of the Eagle Owl (Bubo bubo) used for supportive breeding. Molecular Ecology Notes 2(2):91-93, https://doi.org/10.1046/j.1471-8286.2002.00156.x https://doi.org/10.1046/j.1471-8286.2002.00156.x publication 2002 publication date Isaksson and Tegelstrom 2002 Primers for Bb101, Bb111, Bb126, and Bb42 were screened for variability in Mexican Spotted Owls. Bb101-1, Bb111-1, Bb126-1, and Bb42-1 were redesigned for optimization attempts. Ultimately, Bb101-1 and Bb111-1 were retained as informative markers in this data release (with the "-1" indicating primers were redesigned and provided in the screen table accompanying this data release). Koopman, M.E. Schable, N.A. Glenn, T.C. 2004 journal article Molecular Ecology Notes 4(3):376-378 online Wiley Koopman, M.E., Schable, N.A., Glenn, T.C., 2004. Development and optimization of microsatellite DNA primers for boreal owls (Aegolius funereus). Molecular Ecology Notes 4(3):376-378, https://doi.org/10.1111/j.1471-8286.2004.00658.x https://doi.org/10.1111/j.1471-8286.2004.00658.x publication 2004 publication date Koopman et al. 2004 BOOW18 was screened for variability in Mexican Spotted Owls, was found to give non-specific products, and ultimately, not used. Korfanta, N.M. Schable, N.A. Glenn, T.C. 2002 journal article Molecular Ecology Notes 2(4):584-585 online Wiley Korfanta, N.M., Schable, N.A., Glenn, T.C., 2002. Isolation and characterization of microsatellite DNA primers in burrowing owl (Athene cunicularia). Molecular Ecology Notes 2(4):584-585, https://doi.org/10.1046/j.1471-8286.2002.00326.x https://doi.org/10.1046/j.1471-8286.2002.00326.x publication 2002 publication date Korfanta et al. 2002 Primers for BUOW7, BUOW1, BUOW13, and BUOW16 were screened for variability in Mexican Spotted Owls. BUOW7-1, BUOW1-1, BUOW13-1, and BUOW16-1 were redesigned for optimization attempts. Ultimately, none were retained as informative markers in this data release (with the "-1" indicating primers were redesigned and provided in the screen table accompanying this data release). Proudfoot, G. Honeycutt, R. Douglas Slack, R. 2005 journal article Molecular Ecology Notes 5(1):90-92 online Wiley Proudfoot, G., Honeycutt, R., Douglas Slack, R., 2005. Development and characterization of microsatellite DNA primers for ferruginous pygmy-owls (Glaucidium brasilianum). Molecular Ecology Notes 5(1):90-92, https://doi.org/10.1111/j.1471-8286.2004.00842.x https://doi.org/10.1111/j.1471-8286.2004.00842.x publication 2005 publication date Proudfoot et al. 2005 Primers for Fepo43, Fepo42, and Fepo5 were screened for variability in Mexican Spotted Owls. Fepo43-1 and Fepo5-1 were redesigned for optimization attempts. Ultimately, Fepo5-1 was retained as an informative marker in this data release (with the "-1" indicating primers were redesigned and provided in the screen table accompanying this data release). Thode, A.B. Maltbie, M. Hansen, L.A. Green, L.D. Longmire, J.L. 2002 journal article Molecular Ecology Notes 2(4):446-448 online Wiley Thode, A.B., Maltbie, M., Hansen, L.A., Green, L.D., Longmire, J.L., 2002. Microsatellite markers for the Mexican spotted owl (Strix occidentalis lucida). Molecular Ecology Notes 2(4):446-448, https://doi.org/10.1046/j.1471-8286.2002.00267.x https://doi.org/10.1046/j.1471-8286.2002.00267.x publication 2002 publication date Thode et al. 2002 Primers for 6H8, 8G11, 13D8, and 15A6 were screened for variability in Mexican Spotted Owls. 6H8-1, 8G11-1, 13D8-1, and 15A6-1 were redesigned for optimization attempts. All were retained as informative markers in this data release (with the "-1" indicating primers were redesigned and provided in the screen table accompanying this data release). Redesigned reverse primer for locus 15A6 was not used. Dial, C.R. Talbot, S.L. Sage, G.K. Seidensticker, M.T. Holt, D.W. 2012 journal article Journal of the Yamashina Institute for Ornithology 44(1):1-12 online Yamashina Institute for Ornithology Dial, C.R., Talbot, S.L., Sage, G.K., Seidensticker, M.T., Holt, D.W., 2012. Interspecies amplification of microsatellite markers in great horned owl Bubo virginianus, short-eared owl Asio flammeus, and snowy owl B. scandiacus for studies of population genetics, individual identification, and paternity determination. Journal of the Yamashina Institute for Ornithology 44(1):1-12, https://doi.org/10.3312/jyio.44.1 https://doi.org/10.3312/jyio.44.1 publication 2012 publication date Dial et al. 2012 Redesigned forward and reverse primers for Bb101, Bb111, and reverse primer for Fepo5. Fridolfsson, A.K. Ellegren, H. 2000 journal article Genetics 155(4):1903-1912 online Oxford University Press Fridolfsson, A.K., Ellegren, H., 2000. Molecular evolution of the avian CHD1 genes on the Z and W sex chromosomes. Genetics 155(4):1903-1912, https://doi.org/10.1093/genetics/155.4.1903 https://doi.org/10.1093/genetics/155.4.1903 publication 2000 publication date Fridolfsson and Ellegren 2000 Henke 2005 indicates his primers were based on Boreal Owl (Aegolius funereus) CHD sequences in Fridolfsson and Ellegren 2000 (GenBank AF181824, AF181825). FIELD: Feathers were collected opportunistically near known Mexican Spotted Owl nesting and roosting sites, opportunistically when found away from known roosting/nesting sites and five feathers were collected from a known adult mortality. Feathers were placed in dry envelopes and stored at ambient temperature. Additionally, 10 tissue samples were obtained from preserved mortalities to allow marker screening with higher quality DNA possible from tissue samples. Collection and sampling occurred between 2016 and 2019. These specimens were preserved in Longmire Buffer (Longmire et al. 1988). All sampling was provided by National Park Service, Walnut Canyon National Monument. Unknown LAB: DNA Extraction and quantification (Talbot et al. 2011): DNA extractions from all tissue sources used a salt extraction procedure described in Medrano et al. (1990) and modified as described in Sonsthagen et al. (2004). DNA extractions from feathers used the same protocol, with the following exceptions: dithiotrhreitol (0.1 mg ml−1) was added to the lysis buffer, and 1% glycogen was added to the DNA precipitation step. DNA concentration for tissue samples was quantified using fluorometry, and diluted to 50 ng μl−1 working concentrations for subsequent analyses. Unknown LAB: Except for locus specific modifications noted below, PCR amplifications were carried out in a final volume of 10 μL and contained 50 ng (or 1 uL) genomic DNA, 0.1 mM dNTPs, 0.5 µmole unlabeled primers, 0.1 µmole IRD-labeled primer, 1 μg BSA, 1× PCR buffer [Perkin Elmer Cetus I: 1.5 mM MgCl2, 50 mM KCl, 10 mM Tris/HCl, pH 8.3, 0.01% gelatin (w/v)], and 3 Units GoTaq Flexi DNA polymerase (Promega). PCR reactions began at 94°C for 2 min and continued with 40 cycles each of 94°C for 30 sec; 50°C for 30 sec; 72°C for 1 min followed by a single 30-min extension at 72°C. Exceptions: For locus SneD202w-1 IRD-labeled primer was increased to 1.5 µmole. For loci Fepo5-1 and Bb111-1 IRD-labeled primer was decreased to 0.75 µmole. For loci Bb101-1 IRD-labeled primer was decreased to 0.50 µmole. For loci 8G11-1 and 15A6 1.00 uL of Betaine was added to enhance PCR product. Adding the same amount of betaine to locus 13D8-1 may or may not improve product. For locus SneA127w, thermocycler parameters were shortened to 94°C for 2 min and continued with 40 cycles each of 94°C for 15 sec; 50°C for 15 sec; 72°C for 30 sec; followed by a 30-min extension at 72°C. Primers originally published in Hsu et al. 2003 (for locus Oe128), Hull et al. 2008 (for loci SneD105, SneD202w, and SneA127), Proudfoot et al. 2005 (for locus Fepo5), and Thode et al. 2002 (for loci 6H8 and 8G11) were redesigned and are provided in the table "mexicanSpottedOwl_screen_pierson.csv" included with this data package. Primers originally published in Isaksson and Tegelstroem 2002 (for loci Bb101 and Bb111) were previously redesigned and published in Dial et al. 2012 and are provided in the same table. PCR products were electrophoresed on a 48-well 25-cm 6% polyacrylamide gel on a LI-COR 4200LR automated sequencer (LI-COR, Inc.) (Sonsthagen et al. 2004) Unknown LAB: Henke (2005) P2Owl and P8Owl primers were redesigned (see "mexicanSpottedOwl_screen_pierson.csv" P2MelOwl/P8MelOwl) after sequencing the CHD gene and reducing the size of the targeted marker from 387/393 bp to approximately 210 base pairs to reduce the chance of allelic dropout when using poor quality samples (e.g. feathers, degraded, opportunistic ground sampling). PCR amplifications were carried out in a final volume of 10 μL and contained 50 ng (or 1 uL) genomic DNA, 0.2 mM dNTPs, 0.4 µmole unlabeled primers, 0.03 µmole IRD-labeled primer, 1 μg BSA, 1× PCR buffer [Perkin Elmer Cetus I: 1.5 mM MgCl2, 50 mM KCl, 10 mM Tris/HCl, pH 8.3, 0.01% gelatin (w/v)], and 0.5 Units GoTaq Flexi DNA polymerase (Promega). PCR reactions began at 94°C for 1:30 min and continued with 40 cycles each of 94°C for 30 sec; 48°C for 45 sec; 72°C for 45 sec; followed by a single cycle of 48°C for 1 min; 72°C for 5 min. Unknown Allele sizes were designated by running and scoring a suite of samples against an M13(-29) DNA sequence ladder of known size and including 2 of these scored individuals in 6 lanes of all subsequent gels to size new genotypes. Negative control amplifications (2 each 96-well reaction) were performed for quality control purposes. Scoring was performed using Gene ImagIR 4.05 software (Scanalytics, Inc., Fairfax, Virginia). For sex determination using the P2MelOwl/P8MelOwl primers, samples with 2 bands were designated females and samples with a single band were designated male. Unknown LITERATURE CITED: Longmire, J.L., Lewis, A.K., Brown, N.C., Buckingham, J.M., Clark, L.M., Jones, M.D., Meincke, L.J., Meyne, J., Ratliff, R.L., Ray, F.A., Wagner, R.P., Moyzis, R.K., 1988. Isolation and molecular characterization of a highly polymorphic centromeric tandem repeat in the family Falconidae. Genomics 2(1):14-24, https://doi.org/10.1016/0888-7543(88)90104-8 Medrano, J.F., Aasen, E., Sharrow, L., 1990. DNA extraction from nucleated red blood cells. Biotechniques 8(1):43 Sonsthagen, S.A., Talbot, S.L., White, C.M., 2004. Gene flow and genetic characterization of Northern Goshawks breeding in Utah. Condor 106(4):826-836, https://doi.org/10.1093/condor/106.4.826 Talbot, S.L., Palmer, A.G., Sage, G.K., Sonsthagen, S.A., Swem, T., Brimm, D.J., White, C.J., 2011. Lack of genetic polymorphism among peregrine falcons Falco peregrinus of Fiji. Journal of Avian Biology 42(5):415–428, https://doi.org/10.1111/j.1600-048X.2011.05280.x Unknown Site names describe general sampling areas, they do not represent specific geograhic locations. mexicanSpottedOwl_screen_pierson.csv Table with loci screened to identify informative markers for use in Mexican Spotted Owl individual identification, including screening results and modifications used to collect data presented in the data package. Presented in a Comma Separated Value (CSV) formatted table. Author defined locus Locus names as published in manuscripts first describing markers. Marker names with a dash one (-1) indicate an Alaska Science Center redesign from reported Genbank sequence to attempt to improve amplification (e.g. moving primer to add a GC clamp or to reduce product size). Author defined These are alphanumeric designators assigned by the original researchers who first identified and described each locus. screening_result The screening_result briefly describes results observed when a particular locus was amplified. Author defined used after redesign This indicates a locus was identified as both variable and scorable in initial screenings making it an informative locus. Optimization included redesigning published primers to enhance amplification. Author defined weak variable This indicates the locus appeared to be variable in initial screenings, but ultimately was too weak to be reliably amplified and scored after additional optimization attempts and was dropped from further use. Author defined non-specific This indicates the locus amplified multiple products in initial screenings, but none could be isolated and scored as a microsatellite independent of non-target products and was dropped from further use. Author defined weak This indicates the locus was too weak to be reliably amplified and characterized after additional optimization attempts and was dropped from further use. Author defined monomorphic This indicates the locus amplified a product as expected but was not variable in initial screenings rendering it uninformative and was dropped from further use. Author defined unreliable feather results This indicates the locus appeared to be variable in initial screenings, but ultimately was too weak to be reliably amplified and scored when attempted with DNA extractions from feathers. After additional optimization attempts, it was dropped from further use. Author defined non-specific not variable This indicates the locus amplified multiple products in initial screenings, and while one of the multiple products appeared to be the target, it was monomorphic and uninformative rendering additional optimization unnecessary. This locus was dropped from further use. Author defined used This indicates the locus appeared to be variable in initial screenings using published primer sequences and was retained for use in the study. Author defined non-specific maybe variable This indicates the locus amplified multiple products in initial screenings, and while one of the multiple products appeared to be the target and perhaps variable/informative, it could not be optimized. This locus was dropped from further use. Author defined used; redesign not used This indicates the locus appeared to be variable in initial screenings using published primer sequences and was retained for use in the study. An attempted redesign to enhance PCR product did not improve product, so original published sequences were used. Author defined source This is the publication citation reference of the original published manuscript describing a given locus. The full citations are provided in the "Sources" section of this metadata record. Author defined This is the publication citation reference of the original published manuscript describing a given locus. orig_species This indicates for which species each locus was first described. Author defined This indicates for which species each locus was first described. original_size Published sizes of loci. Author defined The number or number ranges shown indicate approximate sizes in base pairs as reported in original manuscripts for the species developed in. redesigned_size This indicates the size of the locus as used in this study. Blank cells indicate a locus was not retained for this study and not sized for Mexican Spotted Owl (Strix occidentalis lucida). Author defined This indicates the range of observed allele sizes for each locus in Mexican Spotted Owls (Strix occidentalis lucida) using redesigned primers as noted. redesigned_forward_name If forward primer sequences were modified from original published sequences, this indicates their new primer name designation. Blank cells indicate no redesign was attempted. Author defined This alphanumeric designation generally takes the originally published primer name and appends a "-1" indicating a modification. Bb101-1F and Bb111-1F were previously redesigned in our lab as a part of Dial et al. 2012. redesigned_forward_sequence This is the redesigned primer sequence. Blank cells indicate no redesign occurred. Author defined These are the 5' to 3' primer sequences redesigned from original published loci. redesigned_reverse_name If reverse primer sequences were modified from original published sequences, this indicates their new primer name designation. Blank cells indicate no redesign was attempted. Author defined This alphanumeric designation generally takes the originally published primer name and appends a "-1" indicating a modification. Bb101-1R, Bb111-1R and Fepo5-1R were previously redesigned in our lab as a part of Dial et al. 2012. redesigned_reverse_sequence This is the redesigned primer sequence. Blank cells indicate no redesign occurred. Author defined These are the 5' to 3' primer sequences redesigned from original published loci. number_alleles This indicates the number of alleles observed in each locus. Blank cells indicate a locus was not used in this study. Author defined 2 8 mexicanSpottedOwl_microsat_pierson.csv Table with sample ID's, quality if notable, location details, DNA source material, genetically determined sexes, and allele scores (in base pairs) for 1 sexing locus (CHD) and 11 microsatellite loci. Locus names and order of presentation: CHD, 6H8,8G11, 13D8, 15A6, Bb101, Bb111, FEPO5, Oe128, SneA127w, SneD105w, and SneD202w. Presented in a Comma Separated Value (CSV) formatted table. Author defined sample_ID Labels assigned to samples and used to track samples through laboratory analyses. Author defined Alphanumeric label beginning with SPOW for Spotted Owl, followed by 2 letter code indicating a sample from Walnut Canyon National Monument (WC), or a tissue sample from Arizona (AZ) or New Mexico (NM) outside the Monument, followed by a 2 digit number indicating collection site for the WC samples or year for the AZ/NM samples, ending with an individual specimen number. action This indicates any special notes about a particular specimen. Blank cells indicate nothing notable about a specimen. Author defined DROP This indicates the results from this specimen were of such low quality that data are not presented here. Author defined QC This indicates a second DNA extraction of the same feather was completed and provided concordant results. Author defined site This indicates the general locale a specimen was collected from. Author defined Apache-Sitgreaves National Forest This indicates a tissue sample is from Apache-Sitgreaves National Forest. (Generally located near 33.771833 N, -109.354444 W) Author defined Grand Canyon National Park This indicates a tissue sample is from Grand Canyon National Park. (Generally located near 36.040664 N, -112.122737 W) Author defined Coconino National Forest This indicates a tissue sample is from Coconino National Forest. (Generally located near 35.002865 N, -111.686695 W) Author defined Los Alamos This indicates a tissue sample is from Los Alamos, NM. (Generally located near 35.882086 N, -106.287608 W) Author defined Tonto National Forest This indicates a tissue sample is from Tonto National Forest. (Generally located near 33.824647 N, -111.474323 W.) Author defined Kaibab National Forest This indicates a tissue sample is from Kaibab National Forest. (Generally located near 35.966046 N, -112.129298 W) Author defined Walnut Canyon National Monument This indicates a feather or bone was collected opportunistically from a site in Walnut Canyon National Monument. (Generally located near 35.171904 N, -111.509140 W) Author defined state This indicates the state a sampling site is located in. Author defined Arizona This indicates a site is located within the state of Arizona, United States of America. Author defined New Mexico This indicates a site is located within the state of New Mexico, United States of America. Author defined specimen_type This indicates the DNA source material. Author defined muscle DNA was extracted from a muscle sample. Author defined feather- wing DNA was extracted from a wing feather. Author defined feather- breast DNA was extracted from a breast feather. Author defined bone (feather and tendon extracted) DNA was extracted from a feather and a tendon attached to a bone. This specimen was used as quality control since it had sufficient material to extract multiple times. Author defined feather- wing tail DNA was extracted from a wing or tail feather. Author defined feather- body DNA was extracted from a body feather. Author defined sex Sex as determined by CHD locus amplification. Blank cells indicate the CHD amplification was unsuccessful. Author defined M Indicates CHD locus amplification results had only one band indicating this was the homogametic sex, a male. Author defined F Indicates CHD locus amplification results had w bands indicating this was the heterogametic sex, a female. Author defined CHD_allele1 through SneD202w_allele2 These 24 columns represent 1 sexing locus and 11 microsatellite loci. Each locus genotype is represented by two adjacent columns (i.e., allele1 and allele2) each containing one allele score. Genotype numbers were determined by scoring based on allele size relative to an M13 DNA sequence ladder and to samples established as size standards that were run on each gel. Values of AD indicate suspected allelic dropout for that specimen at that locus, FA indicate a suspected false allele for that specimen at that locus and WK indicates result was too weak to assign a genotype with confidence. A blank cell indicates data were not collected at this locus for that sample. Author defined Allele scores (nucleotide base pairs) for each microsatellite locus. U.S. Geological Survey USGS ScienceBase Team Mailing and Physical

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Denver Colorado 80225 USA 1-888-275-8747 sciencebase@usgs.gov The U.S. Geological Survey, Alaska Science Center is the authoritative source of these data, distributed by ScienceBase (a USGS Trusted Digital Repository). Unless otherwise stated, all data, metadata and related materials are considered to satisfy the quality standards relative to the purpose for which the data were collected. Although these data and associated metadata have been reviewed for accuracy and completeness and approved for release by the U.S. Geological Survey, no warranty expressed or implied is made regarding the display or utility of the data for other purposes or on all computer systems, nor shall the act of distribution constitute any such warranty. Any use of trade, firm, or product names is for descriptive purposes only and does not imply endorsement by the U.S. Government. CSV Tabular data in CSV format; FGDC metadata in XML and HTML formats. https://doi.org/10.5066/P9KEWTRX None 20240820 U.S. Geological Survey, Alaska Science Center Mailing and Physical

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